Linear spectral unmixing analysis in single-molecule FRET spectroscopy for fluorophores with large spectral overlap.
Phys Chem Chem Phys
; 26(23): 16561-16566, 2024 Jun 12.
Article
em En
| MEDLINE
| ID: mdl-38832676
ABSTRACT
Fluorescence resonance energy transfer (FRET) is a highly useful tool to investigate biomolecular interactions and dynamics in single-molecule spectroscopy and nanoscopy. However, the use of spectrally overlapping dye pairs results in various artifact signals that prevent accurate determination of FRET values. In this paper, an algorithmic method of spectral unmixing was devised to extract FRET values of spectrally overlapping dye pairs at the single molecule level. Application of this method allows the determination of both the donor-acceptor composition and the FRET efficiency of the samples labelled with spectrally overlapping dye pairs.
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MEDLINE
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En
Ano de publicação:
2024
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Article